tbuckley/cabot-search huggingface dataset:
P037: High breadth whole exome sequencing of circulating tumor DNA identifies novel recurrent genetic alterations in Hodgkin lymphoma
Authors: Laman Mammadova, Jan-Michel Heger, Julia Mattlener, Sophia Sobesky, Melita Cirillo, Paul J. Bröckelmann, Janine Altmüller, Elisabeth Kirst, Stefanie Kreissl, Mia Lohmann, Peter Nürnberg, Andreas Engert, Peter Borchmann, Bastian Von Tresckow, Sven Borchmann
Journal: HemaSphere (2022)

Hodgkin lymphoma (HL) is a B-cell lymphoma with a generally favorable prognosis. However, side effects of aggressive front-line treatment and difficult-to-treat relapsed/refractory disease are still a clinical challenge. A better understanding of the biological diversity of HL at baseline might help to further individualize the treatment of patients and thus ultimately improve outcome. A broad characterization of the genetic drivers of HL is highly important to improve our understanding of the biological diversity of HL. Here, we present the results of our study using whole-exome sequencing of circulating tumor (ct)DNA to broadly characterize the HL oncogenome in 165 patients. After extraction of germline and cell-free DNA, WES was performed, targeting 37 Mb of the human genome. An in-house customized bioinformatic pipeline including twofold error reduction using unique molecular identifiers (UMIs) and digital error suppression was used as previously described by us (Sobesky et. al., Med, 2021). The median ctDNA concentration per patient was 2.26 log haploid genome equivalents per milliliter (log hGE/ml) (range: 1.41–4.18 log hGE/ml) without significant correlation with clinical parameters such as sex, international prognostic score (IPS) or clinical stage. In addition to the detection of several genetic aberrations involved in the pathogenesis of HL (such as B2M, CSF2RB, GNA13, SOCS1), we were able to detect several novel, recurrently mutated potential oncogenes and tumor suppressor genes. Among the most frequently mutated genes not previously described in HL were olfactory receptor family 10 subfamily G member 7 (OR10G7) (11.5%), Apolipoprotein B (APOB) (10.3%) and Filaggrin 2 (FLG2) (9.7%). Furthermore, several previously undescribed copy number variations (CNV) were detected (losses: 13q13.1, 1p36.13a, 6p21.33; gains: 19q13.42, 9p24, 12q12). To decipher the detected genes’ and CNVs’ contribution to the biological diversity of HL, we performed non-negative matrix factorization. Furthermore, neoantigen load was assessed through human-leukocyte antigen (HLA) sequencing. Detailed results will be presented at the meeting. In conclusion, our ctDNA based WES approach provides sufficient sensitivity to both perform genotyping in HL and detecting novel genomic aberrations potentially involved in the pathogenesis of HL.
P067: A pilot of lung cancer screening for survivors of Hodgkin lymphoma
Authors: Rachel Broadbent, Philip Crosbie, John Radford, Christopher J. Armitage, Ben Taylor, Joseph Mercer, Sean Tenant, Kim Linton
Journal: HemaSphere (2022)

Background: Alkylating agents and thoracic radiation put survivors of Hodgkin lymphoma (HL) at excess risk of lung cancer (30-year cumulative incidence 6.4%). Lung cancer screening (LCS) using low-dose CT thorax (LDCT) scans reduces lung cancer mortality in ever smokers in the general population by detecting early-stage lung cancers. We ran an LCS pilot in HL survivors, who are generally ineligible for LCS programmes aimed at the general population. Methods: 218 5+ year survivors of HL treated with mustine or procarbazine and/or thoracic radiation were identified from a follow-up database (ADAPT) hosted at The Christie NHS Foundation Trust and sent a letter of invitation. Participants underwent a baseline LDCT scan. Scans were reported as negative/indeterminate/positive in accordance with lung nodule guidelines. Indeterminate nodules required a further LDCT scan 3-months later. Participants with a positive scan were referred to lung cancer services. Results: Of the 218 invited to the study 54% were female, median age was 53 (range 25–80), median years since treatment 21 (6–45). 12 were ineligible. 127 (59%) expressed interest in participating. The uptake among eligible responders was 83% (102/123). Participation was not influenced by age or gender. Baseline LDCT scan results in 102 participants were: 90 (88.2%) negative, 10 (9.8%) indeterminate, 2 (2.0%) positive. 3-month surveillance LDCT scan results (n=9) were: positive (2), stable (5), with 2 pending. Among 4 participants with positive baseline or 3-month scans, 1 has been diagnosed with early-stage small-cell lung cancer and treated with curative intent, 1 is undergoing tests for presumed lung cancer and 2 are undergoing further surveillance. Coronary artery calcification was detected in 36.3%. Clinically significant incidental findings including emphysema, bronchiectasis, pulmonary inflammation/infection, metastatic breast cancer and vertebral insufficiency fractures were seen in 20 (19.6%). Notably, 3/35 participants who were ever smokers met the age and risk criteria for LCS through the programme aimed at the general population. Conclusion: LDCT scanning protocols tested in the general population are appropriate for use in the HL survivor population in a future targeted LCS programme. The feasibility of such a programme is contingent on developing methods to identify and contact long-term survivors of HL at risk of lung cancer and on investigating and addressing barriers to uptake.
P113: Promising results with anti-PD-1 therapy in primary refractory Hodgkin lymphoma: a single-centre report
Authors: Caterina Zerbi, Tanja Lazic, Roberta Sciarra, Caterina Cristinelli, Alessandro Mazzacane, Federico Carpi, Virginia Valeria Ferretti, Giulia Gambini, Maurizio Bonfichi, Luca Arcaini, Manuel Gotti
Journal: HemaSphere (2022)

Background: Salvage chemotherapy (CHT) followed by autologous stem cell transplant (ASCT) is considered the standard of care for classical Hodgkin lymphoma (cHL) patients who are primary refractory (PrRef) to first line treatment. However, among the few studies based solely on PrRef cHL cases, it has been reported that this strategy leads to a sustained complete response (CR) in almost a half of PrRef patients (pts). Furthermore, Horning et al. reported an overall survival (OS) of 50% at a median follow up of 42 months in a cohort of 29 PrRef pts. Although it is conventionally believed that chemorefractory pts are poor ASCT candidates, recent studies reported promising results in terms of response rate and survival among PrRef cases receiving anti-PD-1 salvage treatment and subsequent ASCT. Methods: We retrospectively collected 9 consecutive PrRef cHL pts who did not respond to salvage CHT and brentuximab-vedotin (BV) and were treated with pembrolizumab monotherapy as fourth-line. Pts defined as PrRef were those who did not achieve a durable (>90 days) CR at the end of first line therapy or those with a Deauville score of 5 (DS 5) at interim PET (PET-2). Results: The majority of pts were male (67%). Systemic symptoms were reported in 56% of cases. Stage at the onset was 2 in 5 cases and 3 and 4 in 2 pts each. An unfavourable prognostic score was seen in 75% of pts. Nodular sclerosis was the predominant histological variant (67%); LMP-1 was positive in 2 out of 7 evaluable cases. The median number of cycles of pembrolizumab administered was 6 (range, 2 to 9). After anti-PD-1 therapy, 75% pts obtained a CR and 25% a partial response (PR). Seven pts underwent ASCT subsequently to pembrolizumab: among these, 87% pts achieved a metabolic CR and 14% a PR at the pre-ASCT PET scan. Disease evaluation after ASCT showed a CR in 100% of cases. After a median follow-up time of 40.3 months (interquartile range: 20.4–54.7 months) from first-line treatment failure, all pts still maintain a CR. Conclusion: If compared to similar series of PrRef cases reported in literature, our experience using pembrolizumab before ASCT resulted in successful outcomes in terms of quality and duration of response in this difficult-to-treat subset of cHL pts.
P025: RADAR: An international phase III, PET response-adapted, randomised trial in progress, comparing ABVD±ISRT with brentuximab vedotin+AVD±ISRT in patients with previously untreated limited-stage classical Hodgkin lymphoma
Authors: John Radford, Toyin Adedayo, Arzhang Ardavan, Sally F. Barrington, Leanne Berkahn, Stephane Chauvie, Laura Clifton-Hadley, Graham P. Collins, Michael Crump, David Cutter, Darren Edwards, Martin Hutchings, Tim Illidge, Amy A. Kirkwood, Kim Linton, Craig H. Moskowitz, Pip Patrick, Beth Phillips, Lois Shepherd, Sanne Tonino, Judith Trotman, Joanna Williams, Nicole Wong Doo
Journal: HemaSphere (2022)

Figure 1: RADAR trial schema Background: Limited-stage classical Hodgkin lymphoma (lsHL), stages I or II without B-symptoms or mediastinal bulk disease, has excellent long-term survival (≥90%) with modern treatment. Commonly used approaches, based on GSHG and EORTC trials, include 2–4 cycles of ABVD (doxorubicin, bleomycin, vinblastine, dacarbazine) plus 20 or 30 Gy radiotherapy (RT), but there are concerns about the impact of using RT in all patients (pts) on the incidence of 2nd cancers and cardiovascular disease. In the UK NCRI RAPID trial, pts who had a -ve PET scan after 3 cycles of ABVD were randomised between RT and no RT; outcomes in the no RT arm were very good but the non-inferiority margin of -7% was not achieved. The RADAR trial is building on the PET-adapted approach of RAPID but intending to make the chemotherapy more effective by replacing bleomycin with the CD30 targeted antibody drug conjugate, brentuximab vedotin (BV), in the BV+AVD regimen. Early phase trials of BV+AVD in lsHL and the phase III ECHELON-1 trial in advanced HL have demonstrated its efficacy and safety. In addition, RT will be integrated on an individualised basis to pts in RADAR who are PET -ve after initial chemotherapy but with larger nodal masses at baseline. Methods: RADAR comprises 2 parallel identical phase III randomised multicentre trials in the UK, Europe, Australia, New Zealand (trial 1; 642 pts), Canada and the USA (trial 2; 400 pts). Eligibility: age 16–69 years, previously untreated stage IA/IIA HL above the diaphragm, any mediastinal mass ≤0.33 of internal thoracic diameter, informed consent. Pts will be randomised to receive either ABVD or BV+AVD. A centrally-reviewed PET-CT scan after cycle 2 (PET2) will determine further treatment (Deauville score (DS) 1–3: total 3 cycles +/- individualised RT; DS 4: total 4 cycles + 30 Gy ISRT; DS 5: treatment failure, subsequent treatment of clinician's choice). 1y endpoint is progression free survival (PFS) and the trial will have 85% power to show an improvement from 90 to 95% PFS at 3 years (HR of 0.49) with 2-sided 5% alpha. 2y endpoints are CMR rate at PET2, event-free survival (progression, death, PET2 DS 4–5), OS, safety and toxicity including cardiovascular disease and second cancers. Exploratory endpoints: predictive value of PET after 1 cycle, prognostic value of baseline metabolic tumour volume. Embedded translational research aims to identify biomarkers predictive of treatment failure. RADAR opened to recruitment in April 2022.
P012: Nodular Lymphocyte-predominant Hodgkin Lymphoma a rare disease with good prognosis: a retrospective multicenter experience
Authors: Francesca Pierdomenico, Raquel Marques, Dulcelena Neves, João. Gaião. Santos, Christopher Saunders, Rita Pinho Peixeiro, Marta Henriques, Ana Tome, Daniela Alves, Margarida Badior, Tathiane Lopes, Sonia Leocadio, Marília Gomes, Alexandra Monteiro, Rita Coutinho, Ana Luisa Pinto, Anabela Neves, João. Paulo Fernandes, Maria Gomes Da Silva
Journal: HemaSphere (2022)

Nodular lymphocyte-predominant Hodgkin lymphoma (NLPHL) is a rare disease. It differs from Classic Hodgkin lymphoma histologically, biologically and clinically, with a more indolent course, a tendency to late relapses and transformation into high-grade B cell lymphomas. There is no consensus on the best treatment approach. The aim of this study is to characterize the clinical characteristics, first-line therapy, outcomes and prognostic factors of NLPHL in our country. We conducted a multicentric, retrospective analysis of patients (pts) with NLPHL diagnosed between 2003 and 2020 in 11 hemato-oncology centers, with a minimum follow-up (FU) of 1y. Overall and complete response rate (ORR and CRR) were defined according to Lugano criteria. Progression-free (PFS) and overall (OS) survival were analyzed using Kaplan-Meier method. Among 140 pts identified, median age at diagnosis was 40y with 69% males, 30% of pts had advanced disease (stage III-IV), 34% ≥3 nodal areas involved, 11% B symptoms and 24% increased LDH. Detailed histological characterization was available in 68 pts, most having the typical variants (A or B). In 19 pts (14%) a “watch and wait” (WW) strategy was chosen; of these, 42% required treatment after a median of 30 mo, while 58% are still under WW at last FU. In the remaining 121 pts, radiotherapy and combined modality treatment (33% and 34% respectively) were used in localized disease, while chemotherapy-only was utilized in 71% of advanced stages, mostly ABVD (42%). Rituximab was associated in 21% of pts, mainly with CHOP and CVP. ORR was 83% and CRR 81%. In localized and advanced disease ORR was 94% and 65% respectively. With a median FU of 65m (95% CI, 55–75) 17 pts relapsed and 8 transformed into high-grade B cell lymphoma. At relapse, salvage chemotherapy with ASCT and chemotherapy with rituximab were equally used. The median number of treatment lines was 1. Five-year PFS and OS were 73.8% (95% CI, 66.4–82.1) and 94.1% (95% CI, 90.0–98.5) respectively. PFS was lower in advanced stages and in pts with ≥3 nodal areas (p<0.001). At last FU 91% of patients were still alive, with 10 deaths (6 lymphoma related). Our study confirms the good outcomes and prolonged survival of NLPHL. Few pts had histological variant characterization, which is increasingly recognized as potentially contributing to better define prognosis and treatment in advanced stages. Treatment strategies should be adequately defined to optimize efficacy and minimize toxicity.
P053: The cGAS-STING anti-tumor immune response pathway as a potential therapeutic target in classical Hodgkin Lymphoma (cHL)
Authors: Ioanna Xagoraris, Konstantina Stathopoulou, Persa Xyderou, Marzia Palma, Elias Drakos, Theodoros P. Vassilakopoulos, Georgios Z. Rassidakis
Journal: HemaSphere (2022)

Background: Cytosolic DNA of exogenous or endogenous origin triggers activation of cyclic GMP-AMP (cGAMP) synthase (cGAS), a cytosolic DNA sensor, that activates innate immune responses through production of the second messenger cGAMP and subsequently activation of the adaptor protein STING. The latter activates TBK1 and IKK kinases that, in turn, activate IRF3 and NF-κB transcription factors, which induce expression of interferons (IFNs), chemokines and cytokines involved in anti-tumor immune responses. The potential role of cGAS-STING pathway in anti-tumor immune responses in cHL remains unknown to date. Methods: STING expression was immunohistochemically analysed in a pilot study group of 32 untreated patients with cHL and available tissue. The in vitro system included 6 cHL cell lines (MDA-V, L1236, L428, L540, HDLM2, KMH2). Gene expression (RNA level) and protein expression and activation (phosphorylation) of cGAS-STING pathway components at baseline and experimental conditions were analysed by quantitative RT-PCR (RT-qPCR) and Western blot, respectively. The cHL cell lines were treated with a STING agonist and TBK1/IKK inhibitor (Amlexanox) alone or in combination with other agents. Silencing of STING gene was performed using transient transfection with specific STING siRNA. The cGAS-STING-associated anti-tumor immune responses were evaluated by assessing the RNA levels of IFN-β, CXCL10, IFN-γ, and a control gene (GAPDH) with RT-qPCR. Results: Using an arbitrary 10% cutoff, STING was positive in the neoplastic Hodgkin & Reed-Sternberg (HRS) cells of cHL in 20 of 32 (63%) patients with a membranous and cytoplasmic pattern. STING expression at the mRNA and protein level was substantially higher in L1236, L428 and HDLM2 compared to other cHL cell lines. Treatment with STING agonist alone stimulated gene expression of IFN-β and/or CXCL10 at a variable level depending on the cell line indicating functional cGAS-STING anti-tumor immune response pathway in cHL. Knocking down STING gene resulted in decreased CXCL10 and type 1 IFN gene expression by cHL cells. Amlexanox treatment also resulted in downregulation of IFN-β or CXCL10 gene expression in vitro. Conclusion: STING agonists and Amlexanox modulate gene expression of type 1 IFNs in cHL with direct therapeutic implications. A large cohort of cHL patients is currently being analysed for the expression and prognostic significance of STING, and the final results will be available at the time of ISHL 2022.
P062: The efficacy and safety of nivolumab 40 mg therapy versus 3 mg/kg in patients with relapsed and refractory classic Hodgkin lymphoma
Authors: Liudmila Fedorova, Kirill Lepik, Natalia Mikhailova, Elena Kondakova, Yaroslava Komarova, Polina Kotselyabina, Evgenia Borzenkova, Vadim Baykov, Ivan Moiseev, Alexander Kulagin
Journal: HemaSphere (2022)

Table 1: Patient’s characteristics Background: The efficacy of nivolumab (Nivo) at a fixed dose of 40 mg in patients with relapsed and refractory classic Hodgkin lymphoma (r/r cHL) was demonstrated in previous studies. Nevertheless, comparison of the efficacy of low doses of PD1 inhibitors with standard dose 3 mg/kg is currently required. Aims: To compare the results of Nivo 40 mg therapy versus standard dosing regimen of 3 mg/kg for patients with r/r cHL. Methods: The Nivo40 trial (NCT03343665) expanded prospective cohort of patients (group 1, n=51) treated with Nivo 40 mg was compared with the retrospective group 2 (n=116) of patients treated with Nivo 3 mg/kg. The response was evaluated every 3 months by PET-CT using LYRIC criteria. Adverse events (AE) were analyzed by NCI CTCAE 4.0.3. Overall response rate (ORR), progression-free survival (PFS), overall survival (OS) were compared between group 1 and 2. During the survival analysis the PFS was censored by the time of additional therapy initiation. Results: Patient’s characteristics are demonstrated in the table 1. Median follow up was 48 (2–60) months in group 1 and 60 (6–70) months in group 2. Median Nivo cycles was 19 (2–49) and 20 (1–32) respectively. The best response to Nivo therapy was detected at 6 (2–24) and 6 (1–27) cycles respectively. ORR was 68% in group 1 and 67% in group 2. The structure of response in group 1 was: complete response (CR) in 40% of patients, partial response (PR) in 28%, stable disease (SD) in 6%, indeterminate response (IR) in 20% and progressive disease (PD) in 6%; in group 2: CR in 34%, PR in 33%, SD in 5%, IR in 20% and PD in 8%. Median OS was not achieved in both groups, 3-year OS was 97,9% and 96,5% respectively (p=0.243). Median PFS was 21,9 months (95%CI: 16,8–26,9) in group 1 and 18,8 months (95%CI: 13,4–24,2) in group 2, 3-year PFS was 23,6% and 27% respectively (p=0.356). Any grade AE were detected in 65% of patients in group 1 and in 79% in group 2 (p=0.068), 3–4 grade AE were detected in 10% and 19% respectively (p=0.151). Additional therapy after Nivo monotherapy was started in 78% of patients after Nivo 40 mg and in 84% after Nivo 3 mg/kg. Allogeneic stem cell transplantation after Nivo therapy was performed in 5 (10%) patients in group 1 and in 26 (22%) patients in group 2. Conclusion: The efficacy of Nivo 40 mg therapy is comparable to the standard dose of 3 mg/kg in patients with r/r cHL. However, a direct comparison of different doses of Nivo in a prospective study is required.
P038: HLA expression patterns of Hodgkin-Reed-Sternberg cells shape a spatially arranged tumor microenvironment in classical Hodgkin lymphoma
Authors: Berit Müller-Meinhard, Sarah Reinke, Nicole Seifert, Andreas Engert, Paul J. Bröckelmann, Michael Altenbuchinger, Wolfram Klapper
Journal: HemaSphere (2022)

Figure 1: Example of an analysis area. a nTME (green square) within a HRSC cluster, dTME (red square) ≥75 μm (yellow circles) away from the nearest HRSC. b Cell detections for CD30 (yellow), CD68 (green), LAG3 (turquoise), FoxP3 (pink) and CD8 (red). Introduction: Classical Hodgkin lymphoma (cHL) differs from other malignancies in its histologic architecture: Few malignant Hodgkin-Reed-Sternberg cells (HRSC) recruit an immune cell-rich tumor microenvironment (TME). Importantly, HRSC frequently show loss of human leukocyte antigen (HLA) expression potentially disturbing T-cell interactions. We aimed to understand how the expression of this key structures of HRSC-T-cell interaction affect the spatial cellular composition of the TME. Material and Methods: 18 lymph node samples of initially diagnosed cHL patients were scored for HLA-I and -II expression. Multiplex immunofluorescence staining for CD30, CD8, CD68, FoxP3 and LAG3 was performed using Akoya Opal Polaris 7-Color Manual IHC Kit. Digital images were analyzed in three equal sized areas near (nTME; including HRSC clusters) and ≥75 μm distant (dTME) to HRSC using QuPath software (Fig. 1). We analyzed cells regarding the expression of one marker, independent of co-expressions. Results: The content of CD68+ cells, CD8+, and LAG3+ T-cells varied in the proximity of HRSC (nTME) dependent of HLA expression of HRSC. HLA-I+ cases showed higher levels of CD68+, CD8+ and LAG3+ cells compared to HLA-I- cases. However, FoxP3+ cell content in nTME was independent of HLA-I expression. We found only slight differences in cellular composition of nTME comparing HLA-II+ with HLA-II- cases. To illustrate spatial gradients, we compared cellular composition of nTME with dTME. Considering all cases, we found higher content of FoxP3+, LAG3+ and CD68+ cells in nTME. Of note, the enrichment of these cell types in nTME was slightly more pronounced dependent on HLA-I rather than HLA-II expression of HRSC. Interestingly, we found higher CD8+ cell content in HLA-I+ cases. This finding is consistent with bulk analyses, which showed HLA-I dependent upregulation of cytotoxic genes (see Seifert et al., ISHL12 submission). Discussion: In line with previous publications, we demonstrate the HRSC niche to be enriched for FoxP3+, LAG3+ cells, and CD68+ macrophages. The enrichment of certain cell types in HRSC proximity seems to be a general feature of cHL independent of HLA expression, which differs in its composition depending on HLA expression of HRSC. We find that enrichment of CD68+, CD8+ and LAG3+ cells is stronger related to HLA-I than HLA-II expression on HRSC. Together with bulk gene expression data we suggest HLA-I expression of HRSC to be a key determinant of TME composition.